Proteomic analysis of fresh and liquid-stored boar spermatozoa

In this study comparative proteomics was used to define changes in the expression of the spermatozoa proteins during liquid storage. Semen from eight boars was analyzed on the day of collection and after liquid preservation at 15–17 °C for three days. Sperm parameters (concentration, motility, morphology, vitality) and percentage of non-capacitated and acrosomal-reacted spermatozoa were determined. Sperm proteins were extracted and separated by two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and proteomic profiles were computationally compared to highlight differentially expressed protein spots that were, in turn, identified by mass spectrometry. The intensities of four spots were significantly different between fresh and liquid stored sperm. Namely: ATP citrate lyase, chaperonin containing T-complex polypeptide 1 (TCP1) subunit ε and probable phospholipid-transporting ATP-ase were over-expressed in liquid stored sperm, whereas cytosolic non-specific dipeptidase was over-expressed in fresh sperm. These differentially expressed proteins could be used as plausible biomarkers for the evaluation of boar semen quality and spermatozoa survival after liquid storage and could help to address problems associated with sperm preservation

Insect protein-based diet as potential risk of allergy in dogs

Before insects can be used widely as an alternative source of dietary protein, their allerginicity should be investigated. Therefore, the aim of our study was to assess the potential adverse reactions of the immune system of dogs against Tenebrio molitor proteins. Dogs sensitised to storage mites T. putrescentiae and A. siro were included. Clinically healthy and clinically allergic dogs were compared. Proteins were extracted from mealworm larvae and their digestibility determined by in vitro incubation with digestive proteases. Mealworm protein extracts and digests were analysed by SDS–PAGE. Canine sera tested for the presence of mite-specific IgEs were used for subsequent Western blotting. LC-MS/MS analysis was used to identify mealworm proteins and their allergenic potential was predicted with the AllermatchTM tool. The binding of canine sera IgEs to mealworm proteins was confirmedhowever, the differences between the two groups of dogs were not significant. Moreover, no clear correlation was found between sensitisation to storage mites and clinical status of the dogs. Altogether, 17 different proteins were identified, including tropomyosin, α-amylase, and Tm-E1a cuticular protein that are known cross-reacting IgE-binding allergens. Our results suggest that dogs allergic to mites may clinically express also the cross-reactivity with mealworm proteins

Retrospective study of bull semen quality — possible correlation with pesticide use?

Decline in semen quality in humans and increased incidence of male reproductive problems could be caused by different factors, including pesticides that could mimic or block the action of endogenous hormones. If the decline in semen quality is real, and environmental chemicals are at least partially responsible for this decline, similar changes should be observed in animals that live in close connection with humans and are exposed to similar levels of pollutants. In the present study, the semen quality of bulls in the last 30 years was examined with respect to the year of birth. Furthermore, semen quality results were correlated to the total pesticide use in a limited geographical area. The results indicate a notable decrease in both ejaculate volume and total number of spermatozoa in ejaculates of bulls born in the late 1970s, while after that (until 2006) there was no obvious downward or upward trend either in ejaculate volume or in the total number of sperm cells. The amount of pesticides released into the environment increased about twofold in the given period, and linear regression analysis revealed a strong and statistically significant correlation between the amount of pesticides used and the total number of spermatozoa in the ejaculate